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KCNK1 / Potassium channel subfamily K member 1 · IHC design guide
Design Immunohistochemistry for KCNK1
Plan KCNK1 chromogenic IHC-P around variable cytoplasmic staining, with high levels in brain and glandular epithelial cells (HPA tissue IHC). Start the catalog antibody at 5 μg/mL and use matched staining controls (datasheet: 5 μg/mL; standard IHC practice).
Evidence assembled Oct 2026 · For research use; verify linked source records and product datasheet before use
KCNK1 Immunohistochemistry Experimental Design Guide
Expected localisation, validated protocols, controls and antibodies — the at-a-glance facts below, then the full design guide.
Must know before staining
| Expected localisation | Variable cytoplasmic staining (HPA tissue IHC) | |
| Staining pattern | Variable cytoplasmic staining; high in brain and glandular cells (HPA tissue IHC) | |
| Antigen retrieval | Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen) | |
| Positive control | Kidney+4 more · see all | |
| Negative control | None in HPA (detected in all 45 tissues); use no-primary + isotype controls |
Important caveats
Reasons your staining may differ from the expected pattern.
| Fixation | Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A04502-1) | |
| Caveat | Antibody staining has low consistency with RNA expression (HPA tissue IHC) | |
| Regulation | Expression varies by tissue (UniProt) | |
| Isoform / epitope | No annotated isoforms; epitope location may matter (UniProt) |
Section 1
Recommended KCNK1 IHC & IF Protocols
The catalog antibody has an IHC-P protocol (datasheet); the published bladder tissue microarray protocol adds an EDTA retrieval condition (PMC11059691).
| Sample | Tissue sections; selected-image fixative not specified (standard IHC workflow) |
| Fixation | Image fixative and duration unreported (datasheet A04502-1); verify before use. |
| Sectioning | 4–5 µm sections on charged slides (standard) |
| Deparaffinisation | Xylene, graded ethanol series to water (standard) |
| Antigen retrieval | Heat-induced epitope retrieval in citrate buffer, pH 6.0, 20 min at 95–98 °C (standard rule: cytoplasmic / membrane antigen) |
| Peroxidase block | 3% H2O2, 10 min, room temperature (standard) |
| Blocking | 10% normal serum of the secondary host, 30 min, room temperature (standard) |
| Primary antibody | Rabbit anti-KCNK1, 5 μg/mL (datasheet A04502-1) |
| Primary incubation | Overnight at 4 °C (standard) |
| Detection | HRP-polymer secondary, DAB chromogen 5–10 min (standard) |
| Counterstain | Hematoxylin, blue, dehydrate and mount (standard) |
| Expected result | KCNK1-positive staining in cells in tubules of kidney (HPA tissue IHC: High). HPA tissue profile: General cytoplasmic expression at variable levels. High levels in brain and glandular epithelial cells. No signal in the no-primary control. |
💡Decision noteStart with citrate pH 6.0 HIER at 95–98 °C for 20 min (page retrieval rule); compare EDTA retrieval (PMC11059691).
Section 2
What Is the Expected KCNK1 Staining Pattern?
KCNK1 is a four-pass membrane channel that can appear at the cell surface and in recycling endosomes (UniProt O00180 topology and subcellular location). In paraffin-section IHC, expect cytoplasmic staining in kidney tubule cells and placental trophoblasts, with variable staining elsewhere (HPA: High in both cell types; general cytoplasmic profile). HPA rates the tissue staining Approved but reports low consistency with RNA expression (HPA: reliability description).
| Strong cytoplasmic stain in kidney tubule cells or placental trophoblasts (HPA: High in both). | This matches the reported IHC pattern. Assess the stained cells and compartment together; intensity alone cannot establish specificity (HPA: general cytoplasmic profile; general IHC practice). |
| Apical membrane accentuation in kidney proximal tubules or bronchial epithelium (UniProt O00180: subcellular location). | These sites are compatible with reported KCNK1 localization. Other kidney tubules may show intracellular staining, so a purely membranous rule would be too narrow (UniProt O00180: kidney localization). |
| Dominant nuclear staining with little cytoplasmic or membrane signal (HPA: cytoplasmic profile; UniProt O00180: subcellular location). | Treat this as discordant with the supplied localization evidence. Review morphology, counterstain and controls before calling the nuclei KCNK1 positive (general IHC practice). |
| Staining in unexpected cells, or a diffuse deposit across tissue compartments (HPA: reported cell-specific pattern). | Cross-reactivity, endogenous detection activity or background may explain it; compare cell types and the no-primary control before assigning target expression (general IHC practice). |
| No signal in kidney tubule cells or placental trophoblasts (HPA: High in both). | Check assay performance before interpreting the section as negative. HPA reports these cells as strong comparators, although its Approved IHC assessment has low RNA consistency (HPA: tissue IHC reliability). |
💡Expected KCNK1 appearanceA convincing positive is cell-associated cytoplasmic staining, potentially with apical accentuation in the appropriate epithelium, strongest in kidney tubules or placental trophoblasts; widespread nuclear or uniform tissue deposit is suspect (HPA: tissue IHC profile and High cells; UniProt O00180: apical localization; general IHC practice).
| Membrane topology and intracellular distribution (UniProt O00180: four transmembrane segments; recycling endosome). | Surface-only scoring could miss intracellular KCNK1. The supplied record does not identify the antibody epitope, so topology alone cannot set an antigen-retrieval condition (UniProt O00180: topology; general IHC practice). |
| Strength of tissue evidence (HPA: IHC Approved; low consistency with RNA expression). | Use the HPA pattern as a comparison, with a specificity caveat. A matching stain supports interpretation but does not independently validate an antibody in a new assay (HPA: reliability description; general IHC practice). |
| Q: What should IF/ICC show? (HPA: subcellular ICC-IF; UniProt O00180: subcellular location). | A: HPA calls vesicles the approved main location; UniProt also reports membranes and recycling endosomes. Interpret IF/ICC on its separate guide page, since these images do not define an IHC-P protocol (HPA: subcellular ICC-IF; UniProt O00180: subcellular location). |
| Situation | Likely cause | Next action |
|---|---|---|
| Kidney tubules and placental trophoblasts are unstained (HPA: High in both). | An IHC run problem is possible; an unstained comparator does not by itself establish absent KCNK1 (general IHC practice). | Check section integrity and the run's positive and negative controls, then review antigen retrieval and antibody conditions for this assay (general IHC practice). |
| The whole section has diffuse chromogen (general IHC practice). | Inadequate blocking or washing, or endogenous detection activity, can obscure cell-associated staining (general IHC practice). | Inspect the no-primary control; adjust blocking, washing or endogenous enzyme quenching as indicated by that control (general IHC practice). |
| Nuclei dominate while cytoplasm is faint (HPA: cytoplasmic profile). | The pattern conflicts with reported localization and may reflect background or an interpretation error (HPA: tissue IHC profile; general IHC practice). | Compare with counterstain and no-primary control; require a plausible cell-associated pattern before scoring positive (general IHC practice). |
| Staining appears mainly in cells outside the reported pattern (HPA: tissue IHC cell types). | Cross-reactivity or endogenous detection activity is possible; HPA's Approved rating does not resolve a new assay's specificity (HPA: reliability; general IHC practice). | Check negative controls and compare a reported positive tissue on the same run before assigning those cells KCNK1 expression (HPA: High kidney tubules and trophoblasts; general IHC practice). |
| Apical-only scoring makes kidney staining seem inconsistent (UniProt O00180: kidney localization). | KCNK1 is reported at proximal-tubule apical membrane and in subapical or cytoplasmic compartments elsewhere in kidney (UniProt O00180: kidney localization). | Record compartment and tubule morphology separately; compare the result with HPA's High tubule-cell staining without assuming every tubule has one pattern (HPA: kidney; UniProt O00180: kidney localization). |
| Brain stain is weaker than kidney stain (HPA: Medium in cerebellar granular layer; High in kidney tubules). | Those reported IHC levels differ, and HPA warns of low consistency between antibody staining and RNA expression (HPA: tissue IHC and reliability description). | Compare like cell populations and the run controls; do not treat relative brightness across organs as a stand-alone specificity test (general IHC practice). |
Experiment evidence
IHC & IF images showing KCNK1 staining
A04502-1 · IHCImmunohistochemistry of KCNK1 in mouse brain tissue with KCNK1 antibody at 5 μg/ml.
A04502-1 · IFImmunofluorescence of KCNK1 in mouse brain tissue with KCNK1 antibody at 20 μg/ml.
HPA · Cerebral cortexHuman Protein Atlas IHC, Cerebral cortex. Source: https://www.proteinatlas.org/ENSG00000135750-KCNK1/tissue/cerebral+cortex#imid_6185952
HPA · CerebellumHuman Protein Atlas IHC, Cerebellum. Source: https://www.proteinatlas.org/ENSG00000135750-KCNK1/tissue/cerebellum#imid_6185955
HPA · ColonHuman Protein Atlas IHC, Colon. Source: https://www.proteinatlas.org/ENSG00000135750-KCNK1/tissue/colon#imid_6185878
HPA · LiverHuman Protein Atlas IHC, Liver. Source: https://www.proteinatlas.org/ENSG00000135750-KCNK1/tissue/liver#imid_6185887
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA016049; green = KCNK1, blue = nucleus, red = microtubules.
HPA · ICC-IF A-431Human Protein Atlas immunofluorescence, A-431, antibody HPA016049; green = KCNK1, blue = nucleus, red = microtubules.
← swipe / scroll for more images →
Real IHC / IF data behind the staining-pattern expectations in this section.
Sample controls for KCNK1 IHC & IF
🧪Run kidney first: cells in tubules should stain (HPA: High in kidney tubular cells). HPA detects KCNK1 in all 45 scored tissues, so use no-primary and isotype controls for the negative; cells outside the stained tubules should be assessed for background without assuming they are KCNK1-negative (HPA: no negative tissue rows).
Positive control tissue: Kidney (Cells in tubules, HPA High)
Negative control tissue: None in HPA: KCNK1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead.
ICC-IF cell lines (HPA subcellular resource): HPA ICC-IF images show KCNK1 in A-431, HeLa, with annotated localisation: Vesicles (approved) (HPA subcellular).
Technical controls: Include a no-primary, secondary-only control, a control matched to the primary antibody’s host species and isotype, and KCNK1 knockout tissue as a biological negative if available (standard IHC practice). In kidney, quench endogenous peroxidase and check background in tubules before scoring chromogenic staining (standard IHC practice).
⚠️Feasibility: No target-specific fixation window or fixation effect is reported in the supplied evidence; the fixative is unreported in the A04502-1 mouse-brain tissue-IHC caption (A04502-1 tissue-IHC caption). Retrieval dependency is also unreported, so compare antigen-retrieval conditions empirically for paraffin sections (standard IHC practice). The supplied evidence does not establish whether frozen sections or IF are easier; kidney tubular background may complicate chromogenic interpretation (standard IHC practice).
HPA tissue IHC evidence for KCNK1
Comprehensive Human Protein Atlas IHC scoring per tissue (reliability: Approved — Low consistency between antibody staining and RNA expression data.). Rows are taken directly from the HPA tissue chart — click any row's HPA link to view the source.
Positive expression · recommended positive controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| Kidney | Cells in tubules | High | Protein (IHC) | HPA → |
| Placenta | Trophoblastic cells | High | Protein (IHC) | HPA → |
| Adrenal gland | Glandular cells | Medium | Protein (IHC) | HPA → |
| Appendix | Glandular cells | Medium | Protein (IHC) | HPA → |
| Breast | Myoepithelial cells | Medium | Protein (IHC) | HPA → |
Undetected expression · recommended negative controls
| Tissue | Cell type | Level | Evidence | Source |
|---|---|---|---|---|
| None in HPA: KCNK1 is detected in all 45 scored tissues. Use a no-primary (secondary-only) and an isotype control instead. | ||||
Section 3
Advanced KCNK1 IHC Tips
Troubleshoot KCNK1 staining in paraffin sections by checking retrieval, compartment, cell type, controls, and how the signal is scored.
What retrieval should I try when KCNK1 staining is weak in paraffin sections?
Use citrate buffer at pH 6.0 for heat-induced antigen retrieval at 95–98 °C for 20 min (page retrieval rule). Keep section thickness, cooling time, and chromogenic detection consistent while comparing retrieved sections with an otherwise matched untreated section (standard IHC practice). The catalog antibody has a mouse brain tissue-IHC image at 5 µg/ml, but its caption does not report retrieval conditions or fixative (selected SKU A04502-1 caption). If staining remains weak, test an alternative retrieval buffer on adjacent sections as a fallback, and judge improvement by cell-specific signal and background together (standard IHC practice).
Could fixation explain weak or uneven KCNK1 staining?
KCNK1-specific fixation sensitivity is unknown because the selected tissue-IHC caption does not state a fixative (selected SKU A04502-1 caption). For paraffin IHC, record fixative, fixation duration, processing, and section age before attributing differences between specimens to KCNK1 biology (standard IHC practice). Compare sections prepared under matched conditions, using the same citrate pH 6.0 retrieval at 95–98 °C for 20 min (page retrieval rule). Include a previously staining section in each run so a change in detection or processing can be recognized, while treating any fixation effect as an observation to validate (standard IHC practice).
Should KCNK1 appear at the membrane or in cytoplasmic puncta?
Assess both membrane-associated and intracellular staining: KCNK1 is annotated at cell membranes, recycling endosomes, and cytoplasmic vesicles (UniProt O00180 subcellular localisation). HPA describes variable general cytoplasmic tissue staining and an approved vesicular IF location, so a diffuse brown signal alone does not establish membrane localisation (HPA tissue IHC; HPA subcellular). In kidney, apical proximal-tubule and subapical staining are described by similarity; distinguish a luminal rim from surrounding cytoplasm at high magnification (UniProt O00180 subcellular localisation). Record the stained cell type and compartment separately, and compare each with a matched negative control before calling a pattern specific (standard IHC practice).
How can an unknown epitope complicate KCNK1 IHC interpretation?
The supplied record lists 0 isoforms and a single 336-aa chain, so an isoform-specific staining explanation is unsupported here (UniProt O00180 processing; isoforms). KCNK1 has 4 transmembrane segments, extracellular loops, and cytoplasmic termini; the antibody epitope is unspecified, so its accessibility after paraffin processing cannot be assigned to either side (UniProt O00180 topology; selected SKU A04502-1 caption). A glycosylation site at residue 95 and phosphoserine at 326 are annotated, but their effects on this antibody's staining are unreported (UniProt O00180 modifications). If two antibody reagents disagree, compare mapped epitopes and appropriate controls before invoking altered protein expression (standard IHC practice).
How should I check an IHC pattern with multiplex IF?
Use IF as a follow-up to the IHC cell and compartment assignment, with a cell-type marker suited to the specimen, such as cytokeratin for an epithelial population (standard IF practice). HPA reports vesicular KCNK1 localisation in ICC/IF images from A-431 and HeLa cells; that observation does not specify the catalog antibody's IF performance (HPA subcellular; selected SKU A04502-1 caption). Choose spectrally separated red or far-red fluorophores after inspecting unstained tissue for autofluorescence, and include single-label controls (standard IF practice). Because the epitope is unmapped, compare gentle permeabilisation with an unpermeabilised condition before interpreting intracellular versus surface signal (UniProt O00180 topology; standard IF practice).
What controls help distinguish KCNK1 staining from chromogenic background?
Run a no-primary control through the same secondary reagent and chromogen steps, and inspect it for endogenous enzyme activity or nonspecific reagent deposition (standard IHC practice). Include a peroxidase-blocking step before DAB detection, then check whether brown signal persists in the no-primary section; these are general chromogenic workflow steps (standard IHC practice). Titrate from the captioned 5 µg/ml concentration on adjacent sections while keeping citrate pH 6.0 retrieval at 95–98 °C for 20 min constant (selected SKU A04502-1 caption; page retrieval rule). Judge background against tissue morphology and expected cell populations, since HPA reports variable cytoplasmic KCNK1 staining across tissues (HPA tissue IHC).
How should I score KCNK1 staining across paraffin sections? ⚠ ANSWER MARKED FOR VERIFICATION
Define the cell population and compartment before scoring, since HPA reports high staining in kidney tubular cells and placenta trophoblastic cells but only medium staining in several other populations (HPA tissue IHC). For cell-based chromogenic scoring, report the percentage of positive cells and an H-score from intensity categories 0–3, with the scoring rules fixed before review (standard IHC practice). For neuropil or punctate signal, measure positive area or object density per mm² within a defined region instead of assigning a cell H-score (standard image analysis practice; HPA tissue IHC). Normalise to the relevant cell count or tissue area, and apply identical thresholds and controls across batches (standard IHC practice).
When is a positive KCNK1 IHC signal credible?
A credible call has reproducible staining in an identified cell population and plausible compartment, with little corresponding signal in the no-primary control (standard IHC practice). Kidney tubular cells show high HPA staining, while KCNK1 is annotated at membranes and intracellular vesicles; interpret the actual compartment at the resolution of the section (HPA tissue IHC; UniProt O00180 subcellular localisation). Question staining confined to cut edges, necrotic areas, unexpected cell types, or sites retaining endogenous enzyme signal after blocking (standard IHC practice). HPA labels its tissue staining Approved but reports low consistency with RNA expression, so corroborate a surprising pattern independently before treating it as KCNK1-specific (HPA tissue IHC).
Boster reagents
Best KCNK1 / Potassium channel subfamily K member 1 IHC Antibodies
A04502-1 has IHC and IF images from mouse brain tissue (catalog image captions) and listed reactivity with human, mouse and rat (catalog reactivity).
Real IHC data
Anti-KCNK1 Antibody
Cat # A04502-1
A04502-1 is listed for IHC-P and IF (catalog applications). Its IHC and IF images both show mouse brain tissue (catalog image captions); the images do not establish performance in human or rat tissue (catalog image captions).
Which to pick: Choose A04502-1 for paraffin-section IHC: IHC-P is listed (catalog applications), and its own IHC caption shows mouse brain tissue stained at 5 μg/mL (A04502-1 IHC image caption); the fixative is unreported (A04502-1 IHC image caption). For IF, A04502-1 has a mouse brain tissue image at 20 μg/mL (A04502-1 IF image caption); ICC validation is unreported (catalog applications and image captions). For cross-species studies, A04502-1 lists human, mouse and rat reactivity (catalog reactivity), while its displayed tissue images are mouse only (catalog image captions); clonality is unreported (catalog clone field).
Each figure is that product's own IHC / IF validation image from its datasheet.
References
- UniProt Consortium. UniProt entry O00180 (KCNK1_HUMAN, Potassium channel subfamily K member 1).
- Human Protein Atlas. KCNK1 tissue IHC expression (reliability: Approved).
- Human Protein Atlas. KCNK1 subcellular location (ICC-IF): Localized to vesicles..
- Human Protein Atlas. KCNK1 antibody validation summary (2 antibodies).
- KCNK1 promotes proliferation and metastasis of breast cancer cells by activating lactate dehydrogenase A (LDHA) and up-regulating H3K18 lactylation. PLoS biology 2024 — PMC11192366.
- Overexpressed KCNK1 regulates potassium channels affecting molecular mechanisms and biological pathways in bladder cancer. European journal of medical research 2024 — PMC11059691.
- Up-regulated expression of two-pore domain K(+) channels, KCNK1 and KCNK2, is involved in the proliferation and migration of pulmonary arterial smooth muscle cells in pulmonary arterial hypertension. Frontiers in cardiovascular medicine 2024 — PMC10894933.
- PubMed PMID:8605869 — UniProt-cited evidence.
- PubMed PMID:9462864 — UniProt-cited evidence.
- PubMed PMID:9362344 — UniProt-cited evidence.
Immunohistochemistry protocol sheet for KCNK1 (Potassium channel subfamily K member 1). Expected localisation Variable cytoplasmic staining (HPA tissue IHC). Antigen retrieval: Citrate pH 6.0 HIER, 95–98 °C, 20 min (rule: cytoplasmic / membrane antigen). Validated IHC antibody: A04502-1 at 5 μg/mL (the product's own IHC validation image is shown). Controls (HPA): positive Kidney (Cells in tubules); negative None in HPA (detected in all 45 tissues); use no-primary + isotype controls. IHC workflow; specimen preparation unspecified. Protocol steps: 1. Sample: Tissue sections; selected-image fixative not specified (standard IHC workflow); 2. Fixation: Image fixative and duration unreported (datasheet A04502-1); verify before use.; 3. Sectioning: 4–5 µm sections on charged slides (standard); 4. Deparaffinisation: Xylene, graded ethanol series to water (standard); 5. Antigen retrieval: Heat-induced epitope retrieval in citrate buffer, pH 6.0 , 20 min at 95–98 °C…; 6. Peroxidase block: 3% H2O2, 10 min , room temperature (standard); 7. Blocking: 10% normal serum of the secondary host, 30 min , room temperature (standard); 8. Primary antibody: Rabbit anti-KCNK1, 5 μg/mL (datasheet A04502-1); 9. Primary incubation: Overnight at 4 °C (standard). Watch for: Fixation: Matched tissue-IHC evidence does not establish the fixation claim. Validate the specimen-specific method before use. (selected-SKU IHC image A04502-1); Caveat: Antibody staining has low consistency with RNA expression (HPA tissue IHC); Isoform / epitope: No annotated isoforms; epitope location may matter (UniProt); Regulation: Expression varies by tissue (UniProt). Evidence: UniProt O00180, HPA tissue IHC · Approved, HPA ICC-IF · vesicles., Datasheet A04502-1, PMC11059691. QR code links to https://www.bosterbio.com/bosterbio-gene-info-cards/KCNK1/immunohistochemistry-ihc-guide.